KR101576610B1 - Composition preventing or treating andropause comprising hippophae rhamnoides l. extract - Google Patents

Abstract

The present invention relates to a composition comprising a Hippophae rhamnoides L. extract for preventing, alleviating or treating andropause syndrome and, more specifically, to a pharmaceutical formulation comprising a Hippophae rhamnoides L. extract for preventing or treating andropause syndrome which alleviates syndrome induced by andropause by increasing testosterone secretion in body and a novel use as a functional food for prevention or treatment.

Description

Technical Field [0001] The present invention relates to a composition for preventing and treating male menopausal syndrome, which contains vitamin A extract,

The present invention relates to a composition for the prevention and treatment of male menopausal syndrome including vitamin-tree extract, and more particularly to a composition for preventing and treating menopausal symptoms of menopausal diseases, The present invention relates to a novel use of an auxiliary food as a natural material.

Women become menopausal and female hormones are rapidly decreased, resulting in several visible changes, which are called menopause. The term "normal menopause" has been used only in women, but in recent years, the concept of male menopausal syndrome has been emerging as a result of a gradual decrease in male hormone production due to aging. The symptoms of real male menopause are similar to those arising from chemical or surgical castration for the treatment of prostate adenocarcinoma. The terms termed "late-onset hypogonadism (LOH)", "age-induced sexual dysfunction syndrome (LOH)" and " testosterone deciency syndrome, TDS). Andropause is a clinical and biochemical syndrome associated with increased age, defined as a typical symptom and a deficiency of serum testosterone levels, which can negatively affect the function of various organs. Symptoms include decreased sexual desire and erectile function, nighttime depression, decreased cognitive ability, decreased fatigue, weight and muscle mass, and atrophy of the skin.

It is argued that the use of male hormone replacement therapy in a manner similar to the use of estrogen replacement therapy in postmenopausal women. According to one study, significant improvements were reported when male menopausal patients were treated with 40 × 2 mg / day of testosterone undecanoate. However, administration of male hormones has also been associated with the risk of adversely affecting liver, lipid conditions, cardiovascular and prostate diseases. In addition, in terms of adenomas and adenocarcinomas, it is known that about 30% of patients with symptoms attributable to male menopause can not receive male hormone replacement therapy. Therefore, there is a need for the development of new therapeutic agents useful in the prevention and treatment of male menopausal syndromes caused by aging, or chemical or surgical castration, which do not provide disadvantages of hormone replacement therapy.

The use of natural product-derived pharmaceutical compositions in the field of drug development is increasing worldwide. Vitamin tree is a bamboo plant, a shrub resembling a willow tree, native to 2-69 degrees north and 27-69 degrees north latitude. The antiviral and anti-inflammatory effects of vitamin-tree-derived flavonoids have been reported.

U.S. Published Patent Application No. 2005-0288325 (December 29, 2005)

 Morales, A. and Lunenfeld, B., Aging Male, 4: 151-162, 2001  Rigatti P., Scattoni V., PSA: Antigene prostatico specifico. Edizioni Medico Scientifiche (EDIMES) Pavia, 1997

It is intended to provide a composition for preventing, ameliorating or treating male menopausal syndrome including vitamin tree extract.

It is another object of the present invention to provide a pharmaceutical preparation and a functional food for preventing, ameliorating or treating male menopausal syndrome which contains vitamin-A extract as an active ingredient.

According to one aspect of the present invention, there is provided a composition for preventing or treating male menopausal syndrome, which comprises vitamin Hippophae rhamnoides L. extract.

In addition, the vitamin tree extract may be extracted from at least one selected from the group consisting of leaves, branches, roots and fruits of a vitamin tree.

In addition, the vitamin tree extract may be an extract soluble in a polar solvent containing an alcohol having 1 to 4 carbon atoms.

In addition, the polar solvent may include a fermentation alcohol.

In addition, the polar solvent may contain a fermented alcohol at a concentration of 20 to 40%.

In addition, the male menopausal syndrome may be one induced by aging.

In addition, the male menopausal syndrome may be one induced by chemical or surgical castration.

According to another aspect of the present invention, there is provided a pharmaceutical preparation for preventing or treating male menopausal syndrome characterized by containing vitamin Hippophae rhamnoides L. extract as an active ingredient.

In addition, the pharmaceutical preparation may further comprise a pharmaceutically acceptable carrier.

According to another aspect of the present invention, there is provided a functional food for preventing or ameliorating male menopausal syndrome, which comprises an extract of vitamin wood ( Hippophae rhamnoides L.) as an active ingredient.

A composition comprising a vitamin A extract according to one aspect of the present invention may increase the level of testosterone in the body. This can be used in the medical and food industries for the purpose of preventing, ameliorating and treating male menopausal disorders and male diseases, as it can alleviate the symptoms caused by menopausal symptoms, including muscle weakness and fatigue. have.

In addition, the composition comprising vitamin extract according to one aspect of the present invention does not induce apoptosis of normal cells and is thus safe for human body, and thus can be used as safe medicines or food additives.

In addition, the composition containing the vitamin tree extract according to one aspect of the present invention can reduce the toxicity and side effects due to the natural plant-derived extract.

FIG. 1 is a graph showing that the vitamin tree extract of the present invention does not show cytotoxicity in TM3 cells,
FIG. 2 is a view showing that the vitamin tree extract of the present invention does not show a protective effect against cytotoxicity induced by hydrogen peroxide (H ₂ O ₂ ) in TM3 cells,
FIG. 3 is a graph showing that the vitamin tree extract of the present invention has an effect of increasing testosterone secretion in TM3 cells,
FIG. 4 is a graph showing that the vitamin tree extract of the present invention has an effect of increasing testosterone secretion upon induction of hydrogen peroxide (H ₂ O ₂ ) in TM3 cells,
FIG. 5 is a graph showing that the vitamin-tree extract of the present invention has an effect of increasing the body testosterone secretion in old rats,
6 is a view showing a rotarod exercise device used for measuring a muscle strength of a rat,
FIG. 7 is a graph showing that the vitamin tree extract of the present invention has an effect of improving muscular strength in old male rats,
8 is a view showing a treadmill exercise device used for measuring the fatigue of a mouse,
FIG. 9 is a graph showing that the vitamin tree extract of the present invention alleviates fatigue in old rats.

The present invention relates to a composition, a pharmaceutical preparation and a functional food for preventing, ameliorating or treating male menopausal symptoms characterized by comprising a vitamin Hippophae rhamnoides L. extract.

Hereinafter, the present invention will be described in more detail.

The composition of the present invention contains vitamin tree extract as an active ingredient.

In the present invention, the part of the vitamin tree which can separate the vitamin tree extract is not particularly limited, and at least one selected from the group consisting of leaves, branches, roots and fruits of a vitamin tree can be used. Preferably, at least one selected from the group consisting of leaves, branches and roots of vitamin trees is selected. More preferably, the leaves of the vitamin tree are selected. This can be cut, dried and pulverized by a conventional method. The drying may be natural drying or freeze-drying.

The method for isolating the vitamin-tree extract is not particularly limited, but a method commonly used in the art can be used for producing the extract. For example, a method using an extraction device such as hot water extraction, immersion extraction, reflux cooling extraction, supercritical extraction, subcritical extraction, high temperature extraction, high pressure extraction and ultrasonic extraction, or a method using an adsorption resin containing XAD and HP-20 Method, but the present invention is not limited thereto. Preferably, it can be obtained by treating with an extraction solvent, extracting it with hot water, and concentrating it under reduced pressure.

As the extraction solvent, a polar solvent may be used. The polar solvent may include, but is not limited to, at least one selected from the group consisting of water, alcohol, acetic acid, dimethyl-formamide (DMFO), and dimethyl sulfoxide (DMSO). Examples of the alcohol include at least one selected from the group consisting of methanol, ethanol, propanol, butanol, n-propanol, iso-propanol, n-butanol, 1-pentanol, 2-butoxyethanol and ethylene glycol But is not limited thereto.

In the present invention, the vitamin tree extract is preferably an extract soluble in a polar solvent containing an alcohol having 1 to 4 carbon atoms. More preferably, it is an extract which is soluble in a polar solvent containing ethanol. The ethanol may be a fermentation alcohol. Fermented alcohol is a vegetable ethanol made by fermenting starch and carbohydrate raw materials with yeast. It is harmless to human body and is used for mainstream, food and beverage, cosmetics and extraction solvent. The ethanol ratio of commonly used fermentation broth is 95% or more. The fermentation alcohol may be diluted in water, and the concentration of the diluted fermentation alcohol is preferably 20 to 40%.

When extracting the extractant by treating the extracting solvent, the extracting solvent may be added at 1 to 100 times the dry weight of the whole vitamin tree. Preferably, it is added at 5 to 20 times the dry weight of the whole vitamin tree.

The temperature at which the extractant is separated by treating the extraction solvent may be 10 to 120 ° C. Preferably 30 to 100 < 0 > C. The vitamin A extract of the present invention can be isolated by extraction for 1 to 72 hours within the above temperature range. Preferably 2 to 12 hours, more preferably 3 to 7 hours, to isolate the vitamin tree extract of the present invention.

The composition comprising the vitamin tree extract of the present invention increases the level of testosterone in the body. This can be used to prevent, ameliorate, or treat male menopausal symptoms caused by decreased secretion of testosterone. In the present invention, male menopausal syndrome includes male-menopausal-induced syndromes, male-menopausal disorders, and male-menopausal-related diseases. Examples include, but are not limited to, mood changes due to hormonal changes, sleep disorders, weight and muscle mass loss, muscle weakness, visceral fat increase, sexual dysfunction, cognitive impairment, fatigue and depression. The male menopausal syndrome may be induced by aging, or by chemical or surgical castration.

The composition comprising the vitamin tree extract of the present invention can be prepared as a pharmaceutical preparation. The pharmaceutical preparations of the present invention can be prepared by using at least one selected from the group consisting of pharmaceutically acceptable carriers, diluents and excipients according to a method which can be easily carried out by those having ordinary skill in the art to which the present invention belongs. Or by formulating it into a unit dose form or by inserting it into a multi-dose container. The formulations may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of excipients, powders, granules, tablets or capsules. The formulation may further comprise a dispersant or stabilizer.

If desired, the pharmaceutical preparation may further comprise a pharmaceutically acceptable carrier. Such pharmaceutically acceptable carriers are those conventionally used in the field of the present invention and include for example lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, But are not limited to, crystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

If desired, the pharmaceutical preparations of the present invention may further contain additives known in the art in addition to the above components. For example, it may further include at least one selected from the group consisting of a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifying agent, a suspending agent, and a preservative agent, but is not limited thereto.

The pharmaceutical preparation of the present invention can be administered orally or parenterally. In case of parenteral administration, it can be administered by skin application, rectal injection, intravenous injection, subcutaneous injection, muscle injection, intraperitoneal injection and transdermal administration. A suitable dosage of the pharmaceutical composition of the present invention may be controlled by such factors as the formulation method, the manner of administration, the age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate and responsiveness of the patient And the ordinarily skilled physician can readily determine and prescribe dosages effective for the desired treatment or prophylaxis. The daily dosage of the pharmaceutical composition of the present invention may be 0.01 mg / kg to 10 g / kg, preferably 1 mg / kg to 1 g / kg.

The composition comprising the vitamin tree extract of the present invention can be produced as a functional food. The functional food is a food prepared by adding a vitamin-tree extract to a food material or a vitamin-tree extract prepared by capsules, powders, suspensions or the like. When the food is taken, the functional food may have a health- Food. Examples of the food include drink, meat, sausage, bread, biscuit, rice cake, chocolate, candy, snack, confection, pizza, noodle, gum, ice cream, soup, beverage, tea, vitamin complex But is not limited thereto.

If necessary, the functional food of the present invention may further comprise components that are ordinarily added in food production. For example, there may be mentioned nutrients, vitamins, minerals (electrolytes), flavors, coloring agents, thickening agents, pectic acid, pectate salts, alginic acid, alginate, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, , Carbonating agent, pulp, and the like, but is not limited thereto.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in more detail with reference to the following examples. It will be apparent to those skilled in the art that these embodiments are illustrative of the present invention and are not intended to limit the scope of the appended claims and that various changes and modifications may be made to the embodiments within the scope and spirit of the invention , And it is natural that such variations and modifications fall within the scope of the appended claims.

Example

≪ Preparation of vitamin tree extract >

Vitamin tree leaves, branches and roots were cut off and dried to remove moisture. A 30% fermented liquor mixed with 80 kg of vitamin wood powder was added in a volume of 10 times by volume and extracted at 100 ° C for 4 hours. After filtration using a filter paper, the supernatant was decompressed and concentrated, followed by lyophilization to obtain a 30% ethanol extract of vitamin wood.

<Cytotoxicity measurement>

Before confirming the effect of the vitamin tree extract of the present invention, a cytotoxicity test was carried out to determine a suitable concentration which does not show toxicity to cells and can induce apoptosis without inducing apoptosis. The cytotoxicity of Vitamin A extract was measured in TM3 cell line derived from mouse testis cells. TM3 cells were cultured in DMEM supplemented with 10% FBS and 1% penicillin / streptomycin at 37 ° C and 5% CO ₂ .

TM3 cells were cultured in a 96-well plate at a concentration of 1 × 10 ⁵ cells / ml at 37 ° C and 5% CO ₂ . Vitamin tree extracts were treated at 1, 3, 10, 30 and 100 ㎍ / ㎖ in each well and incubated for 24 hours. MTT solution was added. After 3 hours, the supernatant was removed. 100 μl of DMSO was added to each well. Absorbance was measured at 540 nm using an ELISA reader. The results are shown in FIG. For reference, Figs. 1 to 5, 7, and all the measurement results shown in Figure 9 is expressed as mean and standard deviation, and statistical significance verification is less than the p-values by using the Student's t-test method 0.05 (p < 0.05) were statistically significant.

Referring to FIG. 1, it can be confirmed that vitamin A extract does not show cytotoxicity against TM3 cells at all concentrations treated.

<Measurement of cytoprotective effect on hydrogen peroxide-induced cytotoxicity>

The cytotoxic effect of hydrogen peroxide (H ₂ O ₂ ) induced by active oxygen species was measured in TM3 cells. TM3 cells were cultured in a 96-well plate at a concentration of 1 × 10 ⁵ cells / ml at 37 ° C and 5% CO ₂ . Vitamin tree extract and hydrogen peroxide were added to each well and cultured for 24 hours. The concentrations of vitamin extracts were 1, 3, 10, 30 and 100 μg / ㎖ per well, and hydrogen peroxide was treated to 200 μM in all wells. MTT solution was added. After 3 hours, the supernatant was removed, and 100 μl of DMSO was added per well. Absorbance was measured at 540 nm using an ELISA reader. Based on the results shown in Fig. 1, cell viability induced by hydrogen peroxide was measured, and the results are shown in Fig.

Referring to FIG. 2, TM3 cells induced by hydrogen peroxide cytotoxicity showed 68.6% viability and significantly decreased cell viability ( ^### p <0.001) as compared with the normal group. Vitamin tree extracts increased the viability of TM3 cells induced by hydrogen peroxide, but did not show any significant difference compared to those without vitamin - tree extract treatment. Through this, it can be confirmed that the vitamin tree extract does not show the cell protection effect against oxidative stress induced by hydrogen peroxide.

&Lt; Measurement of testosterone secretion increasing effect of TM3 cells &

TM3 cells were cultured in a 6-well plate at a concentration of 2 × 10 ⁵ cells / ml at 37 ° C and 5% CO ₂ . Vitamin tree extracts were mixed in FBS-free DMEM at concentrations of 1, 3, 10, 30 and 100 μg / ml, and cultured in each well for 24 hours. Testosterone ELISA kit (ADI-900-065, ENZO Life Sciences, USA) was used to measure the testosterone content. The results are shown in FIG.

Referring to FIG. 3, it can be seen that testosterone levels were significantly increased in a concentration-dependent manner when the vitamin-tree extract was treated. Through this, it can be confirmed that the vitamin tree extract increased testosterone secretion of TM3 cells.

&Lt; Measurement of increase of testosterone secretion of TM3 cells under hydrogen peroxide induction condition >

TM3 cells were cultured in a 24-well plate at a concentration of 2 × 10 ⁵ cells / ml at 37 ° C and 5% CO ₂ . Vitamin tree extract and hydrogen peroxide were mixed in FBS free DMEM medium, and each well was cultured for 24 hours. The concentrations of vitamin extracts were 1, 3, 10, 30 and 100 μg / ㎖ per well, and hydrogen peroxide was treated to 200 μM in all wells. Testosterone ELISA kit (ADI-900-065, ENZO Life Sciences, USA) was used to measure the testosterone content. The results are shown in FIG.

Referring to FIG. 4, the testosterone concentration of TM3 cells induced by hydrogen peroxide cytotoxicity was significantly decreased ( ^# p <0.05) as compared with TM3 cells not cytotoxic. In the case of TM3 cells induced with hydrogen peroxide cytotoxicity, testosterone secretion was significantly increased when treated with vitamin tree extract at the concentrations of 1, 3 and 10 μg / ml.

&Lt; Measurement of increase of secretion of testosterone in body &

The effect of increasing testosterone secretion at the in vivo level was confirmed using aged male rats. Male rats at 6 months of age were matched for 1 week, and were divided into two groups. The experimental animals were kept in a temperature of 23 ± 1 ℃, humidity of 45 ± 5%, 12 hours of light intensity, and water and diet were freely consumed. Vitamin tree extracts were dissolved in distilled water at a concentration of 300 mg / kg, and then administered orally at the same time of day, and the control group was orally administered the same amount of distilled water. Oral administration was performed for 7 days. After the last oral administration, blood was collected from the rat tail and then centrifuged at 4000 rpm for 10 minutes to separate the serum. Testosterone ELISA kit (ADI-900-065, ENZO Life Sciences, U.S.A) was used to measure the testosterone content in the separated serum. The results are shown in FIG.

Referring to FIG. 5, serum testosterone concentration was significantly increased by 83% in the test group (HR300) administered with vitamin extract ( p <0.01). Through this, we can confirm that the vitamin tree extract increased the testosterone secretion in the rat.

<Measurement of muscle strengthening effect>

The muscle strengthening effect of the vitamin extracts was measured by evaluating the muscle strength by exercising the aged male rats using the rotatable cylindrical rotarod shown in Fig. The ages of the rats, the rearing condition and the vitamin-tree extract administration conditions were the same as those of the test for increasing the testosterone secretion in the body, and the rats were forced to walk on the rotary rod at a rotation speed of 20 rpm on the last day of administration, . Twenty-four hours later, the mice were again raised on a cylinder rotating at the same speed, and the time for the mice to lose balance and fall off the cylinder was measured. The measured results are shown in Fig.

As shown in FIG. 7, when the muscle condition was evaluated through the Rota Rod exercise, the Rota Rod exercise time of the test group (HR300) administered with the Vitamin A tree extract was significantly increased compared with the Rota Rod exercise time of the control group, It can be confirmed that the effect of strengthening muscles is demonstrated in this aged male rat.

<Measurement of fatigue improvement effect>

The fatigue improvement effect of the vitamin A tree extract was measured by applying a treadmill exercise device using an electrophoresis stimulation shown in FIG. 8 to old male rats. The rotational speed of the rotating roll of the treadmill exercise device was set to 20 rpm, and the angle was inclined toward the electric stimulating plate at an angle of 20 ° with the ground. The ages of the rats, the rearing condition, and the vitamin tree extract administration conditions were the same as those of the test for increasing the body testosterone secretion, and the adaptive training was performed for 10 minutes on the last day of administration. After 24 hours, the rats were put on a treadmill exercise machine rotating at the same speed, and the anti-fatigue activity of vitamin extracts was measured by setting the exercise time to the limit point at which rats were exhausted and could no longer run. The measured results are shown in Fig.

As shown in FIG. 9, the treadmill exercise time of the test group (HR300) administered with vitamin-tree extract significantly increased ( p <0.05) compared to the treadmill exercise time of the control group, , And vitamin tree extract showed improvement in fatigue in aged male rats.

Claims (10)

Vitamin tree ( Hippophae rhamnoides L.) extract,
A pharmaceutically acceptable carrier, diluent, excipient, dispersant, stabilizer, lubricant, wetting agent, sweetening agent, flavoring agent, emulsifier, suspending agent, preservative, nutrient, vitamin, mineral (electrolyte) A menopausal syndrome consisting of at least one member selected from the group consisting of a colorant, a colorant, a stabilizer, a pectic acid, a pectate, an alginic acid, an alginate, an organic acid, a protective colloid thickener, a pH adjusting agent, a preservative, glycerin, In a pharmaceutical composition for preventing or treating,
Wherein the vitamin tree extract increases testosterone secretion. &Lt; RTI ID = 0.0 &gt; 11. &lt; / RTI &gt;
The pharmaceutical composition for preventing or treating male menopausal symptoms according to claim 1, wherein the vitamin tree extract is extracted from at least one selected from the group consisting of leaves, branches, roots and fruits of a vitamin tree.
The pharmaceutical composition for preventing or treating male menopausal symptoms according to claim 1, wherein the vitamin tree extract is an extract soluble in a polar solvent containing an alcohol having 1 to 4 carbon atoms.
4. The pharmaceutical composition according to claim 3, wherein the polar solvent comprises a fermented alcohol.
5. The pharmaceutical composition according to claim 4, wherein the polar solvent comprises a fermented alcohol at a concentration of 20 to 40%.
The pharmaceutical composition according to claim 1, wherein the male menopausal syndrome is induced by aging.
The pharmaceutical composition according to claim 1, wherein the male menopausal syndrome is induced by chemical or surgical castration.
Vitamin tree ( Hippophae rhamnoides L.) extract,
A medicament for the prophylaxis or treatment of male menopausal syndrome consisting of at least one selected from the group consisting of pharmaceutically acceptable carriers, diluents, excipients, dispersants, stabilizers, lubricants, wetting agents, sweeteners, flavors, emulsifiers, suspending agents and preservatives In a pharmaceutical formulation,
Wherein said vitamin A extract increases testosterone secretion. &Lt; RTI ID = 0.0 &gt; 11. &lt; / RTI &gt;
delete Vitamin tree ( Hippophae rhamnoides L.) extract,
A preservative, a colloid thickener, a pH adjuster, a stabilizer, a preservative, an antiseptic, an antiseptic, an antiseptic, a pectic acid, a pectate, , Glycerin, an alcohol, and a carbonating agent, the functional food for preventing or improving male menopausal syndrome,
Wherein said vitamin tree extract increases testosterone secretion. &Lt; RTI ID = 0.0 &gt; 18. &lt; / RTI &gt;

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